Objective To screen the human pancreatic cancer-associated genes by cDNA microarray.Methods The PCR products of 14 000 genes were spotted onto a chemical-material-coated-glass plates in array.DNAs were fixed onto the glass plate after series of treatments.The total RNAs were isolated from 4 specimens of the pancreatic cancer and the normal tumor-surrounding pancreatic tissues.Both equal quantity RNAs were reversely transcribed to cDNAs and labeled with the fluorescent Cy5-dCTP and Cy3-dCTP to prepare the hybridization probes.The mixed probes were hybridized to the cDNA microarray.After high-stringent washing,the fluorescent signals were scanned by ScanArray 4000 scanner.The values of Cy5-dCTP and Cy3-dCTP on each spot were analyzed and calculated by GenePix Pro 3.0 software.Results By applying this cDNA microarray,189 differentially expressed genes in pancreatic cancer,whose ratios of Cy5/Cy3 were higher than 2.0 or lower than 0.5,were screened out among the 14 000 target genes,comprising 101 known genes and 88 novel gene.Among the known genes,upregulated and downregulated genes were 50 and 51 respectively,including oncogenes and tumor suppression genes,cell cycle related genes,cell skeleton and cinetc-protein related genes,cell apoptosis related genes,DNA synthesis related genes,DNA damage and repair-related genes,transcription factors,receptor related genes,immune related genes,signal transduction related genes,protein translation and synthesis related genes,metabolism related genes.Conclusion The analysis of gene expression profile of tumor based on cDNA microarray can realize high-throughput screening of the genes associated with the pancreatic cancer,and it can help to rapidly explore the genes function.Further analysis of the obtained genes will help to understand the molecular mechanism of the pancreatic cancer.

Objective To identify a novel full-length human pancreatic cancer-associated gene screened by cDNA microarray. Methods One novel full-length gene which overexpressed in the pancreatic cancer was sequenced and analyzed for bioinformatics. The expression of the novel gene in 12 specimens of pancreatic cancer and normal tumor-surrounding pancreatic tissues and 4 pancreatic cancer cell lines were detected by RT-PCR and Northern blot analysis. Results Bioinformatics analysis showed that the chromatosome of the novel gene localized to 4p15, and had an open frame of 501 base pair encoding 166 amino acids of protein. Its theoretical molecular weight was 18 293.23 and PI was 8.57. This protein had more than 90% homologous human hypothetical protein FLJ90013 using BLASTp analysis, and may be a new member of FLJ90013 protein family. The novel gene expressed in 12 specimens of pancreatic cancer tissues and 4 pancreatic cancer cell lines, but not in normal pancreatic tissues. This result was just the same as the result of Northern blot analysis. Conclusion The analysis of gene expression profile of tumor based on cDNA microarray can realize high-throughput screening of the genes associated with the pancreatic cancer, and it can help to rapidly explore the gene function. The novel gene is associated with oncogenesis and development of pancreatic cancer, and might be potential tumor marker or molecular target for diagnosis and treatment of pancreatic cancer.

OBJECTIVETo explore the clinical effect of L-ornithine L-aspartate (LOLA) granules in treating chronic liver disease in patients with high-level serum gamma-glutamyltransferase (G-GT) using a 24-week treatment course.

METHODSTwo-hundred patients with chronic liver disease and above normal G-GT were given a 12-week course of LOLA granules (9 g/d) and then classified into the following three groups according to the change in serum Gamma-GT:group I:patients with Gamma-GT level returned to normal;group II:patients with serum Gamma-GT level that was reduced during the treatment; group III:patients with serum Gamma-GT level that did not decrease or that increased to a higher level than at start of treatment.After the 12-week treatment course, the patients in group I were divided into three subgroups for receipt of a control drug (compound glycyrrhizin, 50mg/d) or an additional 12-week course of Gamma-GT at a reduced dose (LOLA granules 3 g/d) or at the original dose; groups II and III were maintained on the initial dose for an additional 12 weeks.The groups were reassessed at the end of the second 12-week course (at the end of week 24 of the study's observation period).Count data were compared using the x2 test and measurement data were compared using the t-test.

RESULTSIn group I, the serum Gamma-GT level was 90.9% at the end of the first 12-week course and dropped to a mean level of 52.2% for both of the subgroups that received the reduced and original dose after the additional 12 weeks of LOLA granules treatment; the difference from week 12 to week 24 was significant (x2=8.213, P less than 0.05).The 24-week change in serum Gamma-GT levels for the group I reduced and original dose subgroups vs.the control subgroup were also significantly different from those seen in groups II and III (P less than 0.05).The percentage of patients in group I who achieved normal level serum Gamma-GT after 24 weeks of treatment (78.6%) was significantly higher than that for the control group (vs.55.0%, x2=11.452, P less than 0.05).When the patients in group 1 who had received the 12 additional weeks of LOLA granules treatment were measured again at two weeks after the treatments had been discontinued (end of week 26), the percentage of patients with normal serum Gamma-GT level was 92.7%, with only three cases showing obviously abnormal levels; in contrast, the group I patients in the control group of the second 12-week study period had on 66.7% of patients with normal-level serum Gamma-GT.The difference in change between the treated groups (both reduced and original dose) and the control group was significant (x2=14.964, P less than 0.05).

CONCLUSIONPatients whose serumGamma-GT levels returned to normal after receipt of LOLA granules for 12 weeks benefitted from an additional 12 weeks of consolidation treatment, and those given the treatment at the original dose benefitted most.Compared with the compound glycyrrhizin, LOLA granules provided a better maintenance of resolved Gamma-GT level.Therefore, the effect of LOLA appears to be reliable and stable as well as safe for clinical use.

Chronic Disease ; Dipeptides ; therapeutic use ; Humans ; Liver Diseases ; drug therapy ; Liver Function Tests ; gamma-Glutamyltransferase ; blood

ObjectiveTo explore the role and molecular mechanism of Weifuchun （WFC） in inhibiting inflammation and alleviating gastric precancerous lesions （GPL）. MethodHuman gastric mucosal epithelial cells （GES-1） were stimulated with N-methyl-N′-nitro-N-nitrosoguanidine （MNNG） for the modeling of GPL （MC cells）， with Caspase-3 inhibition by Z-DEVD-FMK. MC cells were divided into control （20% blank serum）， WFC （15% and 20% WFC-containing serum）， and caspase-3 inhibitor groups. The cell counting kit-8 （CCK-8） was used to examine the viability of GES-1 cells or MC cells. The Transwell assay and 5-acetylidene-2′-deoxyuridine （EdU） staining were employed to examine cell invasion and proliferation， respectively. Flow cytometry was employed to determine the level of reactive oxygen species. Real-time PCR was conducted to determine the mRNA levels of interleukin （IL）-1， IL-6， and tumor necrosis factor （TNF）-α. Gene Expression Omnibus （GEO） was used to analyze the role of pyroptosis in gastric cancer progression. Western blotting was employed to determine the protein levels of nuclear factor-κB （NF-κB） p65， gasdermin E （GSDME）， and Caspase-3. Immunofluorescence staining was employed to detect the NF-κB p65 protein level and nuclear translocation. Hematoxylin-eosin staining was carried out to observe the pathological changes in the gastric mucosa before and after WFC treatment in the patients. ResultCompared with the control group， MC cells presented enhanced proliferation and invasion energy （P<0.01）. Compared with the blank serum group， WFC-containing serum inhibited the proliferation and invasion of MC cells （P<0.01）， down-regulated the mRNA levels of IL-1， IL-6， and TNF-α， and lowered the level of reactive oxygen species （P<0.05， P<0.01）. The transcriptome data at different stages of gastric cancer showed that pyroptosis was involved in gastric cancer progression， and the GSDME level was significantly higher in GPL patients than in the normal group. Compared with the blank serum， WFC-containing serum lowered the level of NF-κB and inhibited the nuclear translocation of NF-κB （P<0.05）， and it inhibited pyroptosis by suppressing the cleavage of Caspase-3 on GSDME （P<0.05， P<0.01）. The analysis of patient specimens further demonstrated that WFC treatment down-regulated the NF-κB level and GSDME cleavage （P<0.01）， inhibited pyroptosis， and alleviated gastric mucosal inflammation and intestinal epithelial metaplasia. ConclusionPyroptosis is involved in the progression of gastric cancer， and WFC inhibits pyroptosis via the NF-κB/GSDME pathway， thereby alleviating gastric mucosal inflammation in GPL.